GSM7090065: exosomes of EBV-negative cancer cell [AGS_Exo-1_R1]; Homo sapiens; miRNA-Seq
Total RNA was extracted from cells and exosomes using a high purity RNA extraction kit (TRIzol) according to the manufacturer's protocol. The OD260 and OD280 values and ratios of 1 μl extracted samples were determined by ultraviolet spectrophotometer to determine the concentration and purity of extracted samples. The miRNA library was prepared using an RNA sample preparation kit (Epicenter, San Diego, CA, USA) according to the manufacturer's instructions. As outlined below, small RNAs were attached to 3 and 5 splices for reverse transcription to produce single-stranded cDNA. Then PCR amplification was performed. The constructed library was tested for quality and yield using the Agilent 2100 bioanalyzer and ABI StepOnePlus real-time fluorescence quantitative PCR system. Four small RNA libraries were constructed from RNA samples extracted from EBV-positive gastric cancer cell lines (SNU-719) and EBV-negative gastric cancer cell lines (AGS) and their exosomes. The library was sequenced on Illumina HiSeq 2500 platform.